Search Results (551)

Chimeric antigen receptor T (CAR-T) cell therapy has transformed the treatment of hematologic malignancies, yet its broader application, particularly in solid tumors, remains constrained by high cost, labor-intensive manufacturing, limited production capacity, and variable clinical performance, as well as barriers such as poor trafficking, antigen heterogeneity, and an immunosuppressive tumor microenvironment. In vivo CAR-T cell engineering, in which CAR-T cells are generated directly within the patient, offers a paradigm shift by eliminating the need for ex vivo cell processing and complex logistical infrastructure. Among emerging approaches, messenger RNA (mRNA)-loaded lipid nanoparticles (LNPs) have emerged as a promising and clinically tractable platform for in vivo CAR-T cell generation, enabling direct reprogramming of T lymphocytes within the patient and thereby circumventing the need for leukapheresis, viral vector production, and prolonged ex vivo culture, effectively transforming the patient into their own cell therapy factory. This review synthesizes advances in mRNA-LNP-mediated in vivo CAR-T cell generation, encompassing ionizable lipid chemistry and emerging T cell-targeted delivery strategies, including surface functionalization approaches. We discuss the implications of transient CAR expression for immune activation, safety, and therapeutic durability, alongside CAR design optimization through co-stimulatory domains and safety switches. Preclinical evidence from murine tumor models and non-human primates is integrated with current regulatory considerations, and key barriers to clinical translation are highlighted. Collectively, progress in nucleic acid delivery, synthetic immunology, and precision medicine positions in vivo mRNA-CAR-T therapy as a promising modality for oncology and beyond. Full article

Lipid nanoparticles (LNPs) have emerged as popular nucleic acid delivery systems, yet the dynamic mechanisms related to their self-assembly and structural maturation remain insufficiently understood due to the limitations of traditional offline characterization tools. This study establishes a time-resolved (TR) in situ small-angle X-ray scattering (SAXS) methodology to monitor the structural evolution of LNPs during microfluidic formulation and subsequent maturation. By integrating a dual-channel microfluidic mixing system with a SAXS measurement platform, we successfully captured the real-time scattering profiles of both empty and messenger RNA-loaded nanoparticles (mRNA-LNPs). The results demonstrate distinct assembly pathways for empty-LNPs and those encapsulated with mRNA. The empty-LNPs undergo a gradual transition toward periodic nanostructures, whereas mRNA-LNPs exhibit rapid complexation into stable subunits followed by hierarchical assembly. Furthermore, the platform effectively tracked nanoscale structural rearrangements during a microfluidic dilution process, revealed by subtle shifts in scattering peaks and internal periodicity. Overall, this time-resolved approach provides a robust experimental framework for capturing transient intermediate states, offering a valuable tool to elucidate molecular assembly mechanisms and facilitate the rational design of next-generation nanomedicines. Full article

This study reports the green synthesis of silver nanoparticles (AgNPs) using Salvia tomentosa L. leaf extract, and evaluates their physicochemical characteristics and biointerfacial performance, including DNA interaction, antibacterial activity, and antioxidant capacity. AgNP formation was confirmed by UV-Vis spectroscopy through a surface plasmon resonance band at 472 nm. SEM imaging showed predominantly spherical particles with sizes of 30–80 nm and a zeta potential of −17.3 mV, and EDX verified the elemental presence of silver. FTIR spectra indicated that plant-derived biomolecules, particularly phenolics, contributed to the reduction and capping/stabilization of AgNPs. XRD analysis confirmed a crystalline face-centered cubic structure. The AgNPs exhibited moderate, spontaneous binding to DNA (Kb ≈ 1.07 × 10⁴ M⁻¹), characterized by pronounced hyperchromism without evidence of intercalation. Competitive fluorescence assays supported a predominantly non-intercalative, surface-associated interaction with minor groove perturbation, while agarose gel electrophoresis indicated preserved plasmid integrity and no extensive strand cleavage. Collectively, these results suggest reversible and structurally non-destructive AgNP–DNA complexation, indicating their potential for nucleic acid-related nano-biointerface studies, while further investigations are required to evaluate their suitability for biomedical applications. The biosynthesized AgNPs showed enhanced antibacterial activity against Gram-positive (Bacillus cereus) and Gram-negative (Pantoea agglomerans) bacteria compared with the leaf extract, whereas AgNO₃ produced the strongest immediate effect, consistent with rapid Ag⁺ release. Antioxidant activity assessed by DPPH and ABTS assays showed strong radical-scavenging activity for the extract, in line with its high total phenolic content (206.2 mg GAE/g). Although AgNPs displayed lower phenolic content (164.2 mg GAE/g) and reduced antioxidant activity than the extract, they retained moderate scavenging capacity, indicating effective surface functionalization by phytochemicals. Overall, S. tomentosa leaf extract-capped AgNPs combine defined physicochemical features with non-destructive DNA association and antibacterial efficacy, underscoring their promise as phytochemical-functionalized nano-biointerfaces for antimicrobial and related biointerface applications. Full article

Extracellular vesicles (EVs) have attracted growing attention for their diagnostic and prognostic potential as they carry molecular cargo such as DNA, RNA, proteins and lipids derived from their cells of origin. While EV research has traditionally focused on blood, this study explicitly explored saliva as a promising, non-invasive sample matrix for EV isolation and biomarker discovery. Six different EV isolation methods were compared for their ability to recover salivary small EVs suitable for downstream DNA and microRNA analysis. Nanoparticle tracking analysis (NTA) revealed variation in vesicle sizes, concentrations and surface charges across all tested EV isolation approaches. In addition to being the fastest and simplest isolation method, the miRCURY Exosome Isolation kit—serum and plasma from Qiagen (ExiQ) also resulted in the highest EV yields with average particle sizes of ~130 nm. Western blot analysis further verified the presence of EV-specific markers (CD9, Alix) and no detectable signal for ApoA1 as an indicator for lipoprotein contamination, underscoring the purity of ExiQ-isolated vesicles. Always applying the same protocol for parallel DNA and RNA isolation on vesicles extracted by various methods, differences in DNA and RNA yields were observed across the evaluated isolation kits. ExiQ-isolated EVs showed the best recovery for both nucleic acid types. Notably, nuclease treatment of isolated EVs revealed that substantial amounts of DNA were present on the EV surface, whereas microRNA was predominantly localized within the vesicles. The present study, extensively comparing different EV isolation methods, demonstrates that salivary EVs are a viable source for non-invasive diagnostics and suggests the miRCURY Exosome Isolation kit—serum and plasma from Qiagen (ExiQ) to be a good choice for integration in future salivary EV-based diagnostic assays given its simplicity, speed and excellent performance. Full article

Background/Objectives: Lipid nanoparticles (LNPs) are actively being studied as therapeutics and vaccines for various diseases. While LNPs can deliver nucleic acids, their efficiency is limited by the multi-step pathways involved in intracellular trafficking. Crucially, endosomal recycling-driven exocytosis acts as a major problem, rerouting LNPs away from the cytosol and thereby preventing efficient nucleic acid release. Upon entering the cell, LNPs are frequently expelled via endosomal recycling before delivering nucleic acids to cytosol. Previous studies reported that inhibition or deletion of Exo70, a component of the exocyst complex, leads to the accumulation of endosomes because of preventing endosomal recycling. In this study, we investigate the impact of Exo70 inhibition by endosidin-2 (ES-2), an Exo70 inhibitor, on LNP delivery efficiency. Methods: SM-102, cholesterol, DMG-PEG, and DSPC were dissolved in ethanol, while mRNA was dissolved in an aqueous phase to formulate LNPs. Co-treatment of ES-2 with LNPs was performed to evaluate its effect on mRNA delivery, and the resulting delivery efficiency was assessed both in vitro and in vivo. Results: Co-treatment of ES-2 with LNPs significantly enhanced mRNA delivery efficiency, resulting in up to a 4.06-fold increase in vitro and a 3.63-fold increase in vivo. Conclusions: Our findings demonstrate that suppression of Exo70 significantly enhances the mRNA delivery efficiency of LNPs, and this strategy could be applied for the development of mRNA therapeutics. Full article

Chronic liver disease remains a major global health burden, with liver transplantation as the only definitive therapy despite severe limitations in donor availability, surgical morbidity, and patient eligibility. Although the liver has substantial intrinsic regenerative capacity, endogenous repair is often insufficient in chronic injury, cirrhosis, and acute-on-chronic liver failure. As a result, regenerative strategies that restore liver function without whole-organ replacement are increasingly pursued. This review examines controlled release biomaterial-based liver regeneration platforms, particularly those that utilize hydrogels and/or complementary nanoparticle systems, as clinically practical tools to enhance endogenous regeneration. We include discussion of both 3D scaffold-based and injectable hydrogels to enhance regeneration. Used as biological support and controlled release mixtures, they enable local retention, entrapping and controlling the release of regenerative cues including growth factors (HGF, EGF, etc.), nucleic acids for gene expression, stem cells or other cell populations, and conditioned extracellular vesicles, overcoming poor cell engraftment, short cytokine half-lives, and other limitations. Further, synthetic nanoparticles can structure release at the protein/molecular level as well as catalytically modulating oxidative stress and inflammation. Within the context of these systems, we structure the anatomical, engineering, and imaging considerations essential for the clinical translation of gel composite systems while highlighting remaining barriers to wider clinical adoption. Collectively, these advances position biomaterial-enabled regenerative therapies as a realistic alternative or bridge to donor restricted liver transplantation. Full article

The lung represents a promising yet underexploited target for RNA therapeutics due to its large surface area and accessibility via non-invasive inhalation delivery. Despite rapid advances in RNA-based modalities, including small interfering RNA (siRNA), microRNA (miRNA), messenger RNA (mRNA), and CRISPR-Cas systems, efficient pulmonary delivery remains a major challenge. Multiple biological barriers, such as mucus and surfactant layers, mucociliary clearance, immune surveillance, and limited cellular uptake of negatively charged nucleic acids, significantly restrict therapeutic efficacy. In addition, aerosolization processes may introduce mechanical stress, compromising RNA integrity. Nanoparticle-based delivery systems have emerged as a central strategy to address these limitations. By protecting RNA cargo, enhancing mucus penetration, and promoting cellular internalization, engineered nanoparticles enable more effective pulmonary delivery. In this review, we adopt a barrier-centered perspective to examine the key biological obstacles to lung-targeted RNA delivery and highlight recent advances in nanoparticle-mediated strategies, with a focus on lipid nanoparticles, polymeric systems, and inorganic nanomaterials. We further discuss design principles that govern RNA stability, transport, and intracellular release and critically compare the strengths, limitations, and translational potential of each platform, including considerations of toxicity, biodegradability, and clinical readiness. Finally, we outline emerging clinical applications of RNA-loaded nanoparticles, using lung cancer as a representative disease model, and discuss remaining challenges and future directions. Continued innovation in nanoparticle engineering and delivery strategies is expected to accelerate the clinical translation of RNA therapeutics for pulmonary diseases. Full article

Liver cancer remains a significant global health burden, requiring the development of precise nucleic acid delivery systems. Lipid nanoparticles (LNPs) are leading candidates; however, their efficiency is governed by the pK_a of ionizable lipids, which dictates nanoparticle stability and endosomal escape. In this study, we employed a machine learning–driven quantitative structure–activity relationship framework to predict the pK_a of ionizable lipids derived from the DLin–KC2–DMA scaffold. Utilizing a dataset of 56 compounds, we compared Random Forest, Artificial Neural Network, and Extreme Gradient Boosting (XGB) models integrated with Permutation Importance (PI) for feature selection. The optimized PI–XGB model exhibited exceptional predictive accuracy (R² = 0.970, R²_CV = 0.901, RMSE_test = 0.115) and robust generalization confirmed via external validation (RMSE_ext. = 0.313). Mechanistic insights derived from SHapley Additive exPlanation analysis identified charge distribution, molecular topology, and polarity as critical determinants of lipid ionization. These results demonstrate the power of interpretable machine learning in elucidating molecular structure–property relationships, offering a robust computational strategy for the rational design of next–generation ionizable lipids to optimize LNP–mediated gene therapy for liver cancer. Full article

Toxoplasma gondii is an obligate intracellular protozoan parasite that causes toxoplasmosis, posing a significant threat to human health and livestock production worldwide. Although monovalent DNA or mRNA vaccines often confer only partial protection, whether these platforms can be effectively integrated into a heterologous prime–boost regimen against T. gondii remains to be fully elucidated. Here, we constructed GRA35-encoding DNA and mRNA vaccines and evaluated their immunogenicity and protective efficacy, administered either alone or in heterologous prime–boost combinations, in C57BL/6 and BALB/c mice. Both vaccines induced strong antigen-specific immune responses, with the heterologous prime–boost regimen eliciting the strongest effects and conferring the most robust and consistent protection across both mouse strains. Immunization triggered a predominantly Th1-skewed response characterized by significantly elevated IFN-γ production, accompanied by balanced antigen-specific IgG responses. Moreover, vaccinated mice developed rapid and potent cytotoxic T lymphocyte (CTL) responses. Following challenge with the RH and PRU strains, vaccinated mice exhibited prolonged survival and significantly reduced brain cyst burdens following PRU challenge compared with control groups. Collectively, these findings indicate that GRA35-based nucleic acid vaccines, particularly when administered in a heterologous prime–boost regimen, elicit multifaceted protective immune responses and represent promising vaccine candidates against T. gondii infection. Full article

Lipid nanoparticles (LNPs) have become the cornerstone of nucleic acid delivery platforms, particularly in RNA-based vaccines and therapeutics. However, the conventional methods of LNP production, which are primarily reliant on microfluidic mixing of aqueous and organic solvent phases, pose limitations in terms of mRNA stability, residual organic contamination, scalability, cost, and environmental impact. These limitations prompted a renewed search for non-conventional strategies with the promise of improving mRNA-LNP encapsulation approaches. These emerging approaches aim to address key bottlenecks, including mRNA hydrolysis-driven degradation, high production losses, and complex downstream purification. Moreover, the ability to decouple LNP synthesis from mRNA encapsulation could enable streamlined, modular manufacturing workflows and customizable payload delivery, including single- or multiple-mRNA payloads, thereby expanding the therapeutic scope of LNPs. This review offers an early insight into the design principles and scalability potential of emerging non-conventional LNP encapsulation approaches, including solvent-free and microfluidics-free methodologies, and pre-built LNP workflows. We also examine trends in emerging LNP encapsulation tools, including high-shear mixing, sonication, membrane contraction, and other approaches. Finally, we extrapolate the suitability of the methods for scale-up approaches and their economic implications based on the process information. Full article

Cancer remains a major global health challenge, characterized by abnormal cell growth and metastasis. Current limitations of conventional therapies, particularly non-specific toxicity harming healthy cells, highlight the need for more targeted approaches. Nanotechnology offers a revolutionary solution, utilizing nanoparticles (NPs) for precise drug delivery to tumor sites while minimizing off-target effects. These nanometer-scale particles enable superior binding to cancer cell membranes, the tumor microenvironment, or nuclear receptors, facilitating significantly higher local concentrations of therapeutic agents. NPs, synthesized via physical, chemical, or biological methods, are categorized as organic (organic material-based) or inorganic (metallic particle-based). Key delivery mechanisms include the Enhanced Permeability and Retention (EPR) effect and Active Transport and Retention (ATR). This review specifically examines NP applications for the most prevalent cancers in the US (2025): breast, prostate, and lung. Gold and magnetic NPs show significant promise for early breast cancer detection. For lung cancer, polymeric NPs like PCL, PLA, and PLGA are effective carriers for peptides, proteins, and nucleic acids. BIND-014, a docetaxel-loaded NP formulation, represents an emerging strategy for prostate cancer. Clinically established examples include liposomal doxorubicin and albumin-bound paclitaxel. We comprehensively discuss the synthesis methods, delivery mechanisms, and the current landscape of NPs in research and clinical trials for these cancers. This analysis underscores the potential of nanotechnology to provide more effective and targeted therapeutic options for cancer patients in the future. A distinctive feature of this review is its comparative cancer-specific analysis of NP platforms in breast, prostate, and lung cancers. Unlike previous generalized reviews, this work integrates synthesis strategies, delivery mechanisms, translational challenges, and clinically relevant formulations to provide a bench-to-bedside perspective on the future of nanomedicine in oncology. Full article

Plant extracts are widely used as natural pesticides, cosmetic ingredients, and in pharmaceutical applications. However, their poor water solubility and stability limit their usability. Lipid nanoparticles (LNPs) offer an effective encapsulation strategy to overcome these challenges. This study demonstrates the encapsulation of three representative substances from these industries: quercetin as a pesticide, irones as a cosmetic ingredient, and nucleic acids for pharmaceutical use. Ultrasonic treatment was used for the encapsulation of quercetin and irones, and a concept for continuous encapsulation in a plug flow reactor was proposed for process intensification. Inline multi-angle light scattering and dynamic light scattering measurements proved effective for real-time monitoring and enabled the replacement of traditional batch measurements. In the pharmaceutical area, mRNA-based therapies require LNP encapsulation to prevent nucleic acid degradation. Plant-based β-sitosterol was used as an alternative helper lipid to cholesterol, resulting in an average particle diameter of 72 nm and an encapsulation efficiency of 91%, comparable to commercial formulations such as the Comirnaty vaccine. Furthermore, a novel process model based on population balances was developed to simulate the entire manufacturing process, from rapid mixing in a T-mixer to particle stabilization via buffer exchange during diafiltration. By applying a quantitative and distinctive model validation workflow, the model was shown to be as accurate and precise as the experimental data, enabling its use as a digital twin for autonomous continuous operation. In summary, this study contributes to reducing the facility footprint and cost of goods through the implementation of continuous processing and model-based control. This approach improves productivity by 20% and reduces process time by a factor of two. Full article

The utilization of immunomodulatory nanomaterials, i.e., leveraging their unique properties to enhance immune responses, represents a transformative approach for the treatment of various diseases. Recent advancements in nanotechnology have enabled the design of nanomaterials capable of delivering immunomodulatory agents in a targeted manner, such as cytokines, antibodies, and nucleic acids, to specific cells or tissues involved in immune regulation. These nanomaterials, including nanoparticles, liposomes, nanogels, nanoemulsions, dendrimers, MXenes and extracellular vesicles, have been increasingly tailored to modulate immune responses with precision and efficacy. This targeted approach not only enhances therapeutic outcomes but also reduces off-target effects, minimizing systemic toxicity. In this review, an overview of immunomodulatory nanomaterials and their biomedical applications are highlighted. Herein, we have discussed different types of nanomaterials and their design strategies, interactions with different immune system components (macrophages, dendritic cells (DCs), neutrophils, T lymphocytes (CD4⁺ helper T-cells, CD8⁺ cytotoxic T-cells, regulatory T-cells/Tregs, and memory T-cells), and B lymphocytes), and immunomodulation mechanisms. Furthermore, nanomaterial-based immunomodulation strategies to enhance cancer immunotherapy, wound healing, and bone regeneration and the treatment of infectious diseases, autoimmune diseases, and allergy and are discussed in detail. In addition to therapeutic applications, selected nanomaterial platforms demonstrate significant potential in pharmaceutical formulations by improving drug stability, controlled release, and bioavailability, as well as in cosmetology through skin-targeted delivery, anti-inflammatory activity, immune protection, and enhanced tissue regeneration. Finally, clinical trial updates, challenges and future prospects are outlined. Key findings indicate that lipid-based, polymeric, inorganic nanoparticles and dendrimers provide complementary advantages for immunomodulation, including efficient delivery, controlled release, multifunctionality, and precise immune targeting. Despite safety, regulatory, and scalability challenges, these systems show strong potential for advancing precision and personalized medicine. Taken together, these innovations hold great promise for personalized medicine approaches, wherein nanomaterials can be tailored to individual patient profiles for more effective and precise disease treatment and prevention strategies. This review focuses primarily on the mechanistic interactions between immunomodulatory nanomaterials and immune cells, including macrophages, dendritic cells, neutrophils, T lymphocytes, and B lymphocytes, rather than providing an exhaustive treatment of physicochemical optimization parameters such as particle size or surface modification chemistry, which fall outside the defined scope of this work. Full article

Lipid nanoparticles (LNPs) have become an important platform for the delivery of RNA therapeutics, including messenger RNA (mRNA) and small interfering RNA (siRNA). However, most clinically approved LNP formulations exhibit strong liver tropism following systemic administration, which limits efficient delivery to extrahepatic tissues. This inherent biodistribution profile has therefore been recognized as a key challenge for expanding the therapeutic applications of RNA nanomedicine. Recent efforts have focused on engineering functionalized LNP systems to improve delivery specificity beyond the liver. Surface modification with targeting ligands—such as antibodies, peptides, and nucleic acid aptamers—can promote receptor-mediated uptake by specific immune cell populations, including macrophages, dendritic cells and T lymphocytes. In parallel, advances in lipid design have improved intracellular RNA delivery by facilitating endosomal escape. These developments have broadened the potential use of RNA nanomedicine for inflammatory disorders, including autoimmune diseases, neuroinflammation, and cardiovascular inflammation. Functionalized LNPs are also being investigated for in vivo engineering of immune cells. This review summarizes current strategies for designing functionalized LNP systems, highlights their emerging applications in immune and inflammatory diseases, and discusses key challenges for clinical translation. Full article

Colorectal cancer (CRC) remains one of the leading causes of cancer-related mortality worldwide and is frequently diagnosed at an advanced stage due to limitations of current screening methods. Although surgical resection is the standard treatment, conventional tissue biopsies are invasive and restrict real-time assessment of tumor dynamics. Liquid biopsy has emerged as a promising noninvasive approach enabling repeated analysis of tumor-derived components in body fluids. Among these, exosomes have gained considerable attention as potential diagnostic biomarkers in CRC. This review summarizes current evidence on exosome biogenesis, molecular composition, and their diagnostic relevance in colorectal cancer. We discuss exosomal nucleic acids, proteins, and lipids as biomarkers detectable in patient samples, as well as analytical platforms used for their isolation and characterization, including ultracentrifugation-based methods, size-exclusion chromatography, nanoparticle tracking analysis, electron microscopy, proteomics, lipidomics, and sequencing approaches. Accumulating data demonstrate that exosomal microRNAs, long non-coding RNAs, proteins, and lipid signatures correlate with tumor progression, immune modulation, angiogenesis, and epithelial–mesenchymal transition. Advances in microfluidic technologies, Raman/SERS spectroscopy, and AI-based data analysis are contributing to further improvements in diagnostic sensitivity and reproducibility. Despite their potential, the lack of standard isolation and validation protocols remains a major obstacle to clinical implementation, highlighting the need for large-scale multicenter studies before exosome biomarkers can be routinely used in CRC diagnostics. Full article